Genomic sequencing of key genes in mouse pancreatic cancer cells.

TitleGenomic sequencing of key genes in mouse pancreatic cancer cells.
Publication TypeJournal Article
Year of Publication2012
AuthorsWang, Y, Zhang, Y, Yang, J, Ni, X, Liu, S, Li, Z, Hodges, SE, Fisher, WE, Brunicardi, FC, Gibbs, RA, Gingras, M-C, Li, M
JournalCurr Mol Med
Date Published2012 Mar
KeywordsAnimals, Cell Line, Tumor, Cyclin-Dependent Kinase Inhibitor p16, Homeodomain Proteins, Mice, Pancreatic Neoplasms, Polymerase Chain Reaction, Proto-Oncogene Proteins p21(ras), Sequence Analysis, DNA, Smad4 Protein, Trans-Activators, Tumor Suppressor Protein p53

Pancreatic cancer is a multiple genetic disorder with many mutations identified during the progression. Two mouse pancreatic cancer cell lines were established which showed different phenotype in vivo: a non-metastatic cell line, Panc02, and a highly metastatic cell line, Panc02-H7, a derivative of Panc02. In order to investigate whether the genetic mutations of key genes in pancreatic cancer such as KRAS, TP53 (p53), CDKN2A (p16), SMAD4, ZIP4, and PDX-1 contribute to the phenotypic difference of these two mouse pancreatic cancer cells, we sequenced the exonic regions of these key genes in both cell lines and in the normal syngeneic mouse pancreas and compared them with the reference mouse genome sequence. The exons of KRAS, SMAD4, CDKN2A (p16), TP53 (p53), ZIP4, and PDX-1 genes were amplified and the genotype of these genes was determined by Sanger sequencing. The sequences were analyzed with Sequencher software. A mutation in SMAD4 was identified in both cell lines. This homozygote G to T mutation in the first position of codon 174 (GAA) generated a stop codon resulting in the translation of a truncated protein. Further functional analysis indicates that different TGF-β/SMAD signaling pathways were involved in those two mouse cell lines, which may explain the phonotypic difference between the two cells. A single nucleotide polymorphism (SNP) in KRAS gene (TAT to TAC at codon 32) was also identified in the normal pancreas DNA of the syngenic mouse and in both derived tumoral Panc02 and Panc02-H7 cells. No mutation or SNP was found in CDKN2A (p16), TP53 (p53), ZIP4, and PDX-1 genes in these two cell lines. The absence of mutations in genes such as KRAS, TP53, and CDKN2A, which are considered as key genes in the development of human pancreatic cancer suggests that SMAD4 might play a central and decisive role in mouse pancreatic cancer. These results also suggest that other mechanisms are involved in the substantial phenotypic difference between these two mouse pancreatic cancer cell lines. Further studies are warranted to elucidate the molecular pathways that lead to the aggressive metastatic potential of Panc02-H7.

Alternate JournalCurr Mol Med
PubMed ID22208613
PubMed Central IDPMC3799885
Grant ListU54 HG003273 / HG / NHGRI NIH HHS / United States
R21CA133604 / CA / NCI NIH HHS / United States
R01CA138701 / CA / NCI NIH HHS / United States
R21 CA133604 / CA / NCI NIH HHS / United States
R01 CA138701 / CA / NCI NIH HHS / United States

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